The present disclosure provides a protective film for a substrate, a manufacturing method thereof, a substrate and a manufacturing method of a display panel. The substrate includes a plurality of panel regions correspondingly forming a plurality of display panels. Each of the panel regions has a first region corresponding to a bending region of the display panels. The protective film includes a entire film layer and a hollow portion, the size of the entire film layer matches the size of the substrate, and the hollow portion is for exposing the first region in each of the panel regions when the protective film is attached to the substrate.